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Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial
L N Shingarova1, L E Petrovskaya2, A V Zlobinov2,3
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia. lshing@ibch.ru.
Biochemistry. Biokhimiia
|September 10, 2018
Summary
Researchers optimized a bacterial display system to create novel antibody mimetics. They developed new tumor necrosis factor (TNF)-binding proteins using the 10th human fibronectin type III domain (10Fn3) scaffold, showing high affinity for TNF.
Area of Science:
- Biotechnology
- Protein Engineering
- Immunology
Background:
- Antibody mimetics offer a promising alternative to traditional antibodies for medical and biotechnological applications.
- The 10th human fibronectin type III domain (10Fn3) is a versatile scaffold protein for developing these mimetics.
- Optimizing display systems is crucial for efficient screening and selection of functional scaffold proteins.
Purpose of the Study:
- To optimize a bacterial display system for the 10Fn3 scaffold protein.
- To construct novel artificial proteins that bind to tumor necrosis factor (TNF).
- To identify and characterize high-affinity TNF-binding 10Fn3 variants.
Main Methods:
- Construction of a combinatorial 10Fn3 gene library.
- Screening of the library using a bacterial display method.
- Expression and characterization of selected 10Fn3 variants in *Escherichia coli* for TNF-binding activity.
Main Results:
- Successful optimization of the cell display system for the 10Fn3 scaffold.
- Identification of several 10Fn3 variants with high affinity for TNF.
- Characterization of the binding properties of these novel TNF-binding proteins.
Conclusions:
- The optimized bacterial display system is effective for developing 10Fn3-based antibody mimetics.
- Novel high-affinity TNF-binding proteins were successfully engineered using the 10Fn3 scaffold.
- These engineered proteins hold potential for future applications in medicine and biotechnology.
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